Proceedings Article | 5 March 2008
KEYWORDS: Confocal microscopy, Microscopes, Mirrors, Green fluorescent protein, Tissues, Skin, Luminescence, Objectives, In vivo imaging, Sodium
Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively.
CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex
vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential
fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin.
In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The
prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of
some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens,
0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human
volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP
transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and
curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM
imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated
the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has
similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In
vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals
are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by
individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell
border could be seen. In papillary dermis, fluorescein distribution is more homogeneous. Curcumin is a yellow food dye
that has similar fluorescent properties to fluorescein sodium. In vivo CLSM of transgenic GFP mouse enable on in vivo,
high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, skin appendage
and blood vessels. In conclusion, this study demonstrates the usefulness of CLSM as technique for imaging skin in vivo.
In addition, CLSM is non-invasive, the same tissue site may be imaged over a period of time to monitor the various
change such as wound healing, severity of skin diseases and effect of therapeutic management.
